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Research on Determination of 24 Intracellular Metabolites in E.coli by LC-MS/MS
Author(s): 
Pages: 151-156
Year: Issue:  5
Journal: Food and Fermentation Industries

Keyword:  高效液相色谱-串联质谱 大肠杆菌 胞内中间代谢产物 代谢组学;
Abstract: 建立了一种快速、准确的同时测定大肠杆菌(E.coli)24种胞内代谢中间产物的液相色谱-串联质谱分析方法。样品经过淬灭、洗涤和提取后得到胞内代谢物,高速冷冻离心去除杂质。经Waters Atlantis HILIC silica色谱柱分离,以乙腈-10 mmol/L乙酸铵水溶液为流动相进行梯度洗脱,流速为250μL/min。电喷雾负离子(ESI-)模式电离,多反映监测模式(MRM)检测,基质标准溶液法进行定量。结果表明:24种代谢物在0.10~100μmol/L范围内呈良好的线性关系,相关系数均大于0.99;24种代谢物的检出限(S/N>3)为0.05~0.34μmol/L,3个添加水平的回收率为83.7%~108.1%,相对标准偏差(RSD)均小于10%。该方法具有简便、灵敏度高、定性准确等特点。
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